Use of gated perfusion to study early effects of anoxia on cardiac energy metabolism: a new 31P NMR method.
Use of gated perfusion to study early effects of anoxia on cardiac energy metabolism: a new 31P NMR method.
复制标题
使用门控灌注研究缺氧对心脏能量代谢的早期影响：一种新的 31P NMR 方法。
DOI：
10.1021/bi00320a024
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发表时间：
1984
期刊：
影响因子：
2.9
通讯作者：
Chan,SH
中科院分区：
文献类型：
--
作者：
Barbour,RL;Sotak,CH;Levy,GC;Chan,SH
Randall L.克里斯托弗·巴特尔索塔克，乔治C。利维和塞缪尔。P. Chan** 摘要：描述了一种新的31 P NMR方法，该方法能够确定在离体灌注的工作大鼠心脏中各种含磷化合物的细胞内水平的快速变化。该技术涉及对心脏的31 P NMR测量进行门控，所述心脏交替灌注有含有10 mM丙酮酸盐的改良Krebs-Henseleit培养基，并用95%O2/5%CO2或95%N2/5%CO2平衡。实验设计允许在单个O2/N2灌注循环期间进行多达三次NMR测量。当在治疗期间以不同的间隔重复这些测量时，可以确定代谢物水平的快速变化。初步的研究表明，心脏保持血流动力学稳定的有氧/缺氧灌注循环，判断心率，峰值收缩压，主动脉输出量，和冠状动脉流量至少80分钟的磁铁时，受到4.5秒的O2和1.5秒的N2灌注的周期时间。在这些条件下进行的核磁共振测量表明，从完全有氧灌注到该循环的转变揭示了一个新的稳态，无机磷酸盐水平从6%增加到10%，可观察到的总磷含量可能显著降低。检测缺氧对高能磷代谢物水平影响的两种方法是冷冻钳住，然后对中和提取物进行酶分析。（威廉姆森& Corkey，1969）和最近使用的磷-31核磁共振光谱法。前一种技术容易在大多数实验室中进行，但遭受破坏性程序，从而获得
Randall L. Barbour, Christopher H. Sotak, GeorgeC. Levy, and Samuel. P. Chan** abstract: A novel 31P NMR method is described that is capable of determining rapid changes in the intracellular levels of various phosphorus-containing compounds in an isolated, perfused working rat heart. This technique involves the gating of 31P NMR measurements to a heart that is alternately perfused with a modified Krebs-Henseleit medium containing 10 mM pyruvate and equilibrated with either 95% 02/5% C02 or 95% N2/5% C02. The experimental design allows up to three NMR measurements to be made during a single 02/N2 perfusion cycle. When these measurements are repeated at different intervals during thecycle, rapid changes in metabolite levels can be determined. Preliminary studies have shown that hearts remain hemodynamically stable to the aerobic/anoxic perfusion cycle as judged by heart rate, peak systolicpressure, aortic output, and coronaryflow for at least 80 min in the magnet when subjected to cycle times of 4.5-s 02 and 1.5-s N2 perfusions. NMR measurements made under these con-ditions showed that a transition from full aerobic perfusion to this cycle revealed a new steady state, with an increased inorganic phosphate level from 6% total observable phosphorus to 10% and a possibly significant decreased measurement ofTwo approaches to examining the effects of anoxia on the levels of high-energy phosphorus metabolites are freezeclamping followed by enzymatic analysis on neutralized ex-tracts (Williamson & Corkey, 1969) and the more recent use of phosphorus-31 nuclear magnetic resonance spectroscopy. The former technique is easily performed in most laboratories but suffers from being a destructive procedure, thereby ob-